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DNA Research 2001 8(1):33-45; doi:10.1093/dnares/8.1.33
© 2001 by Kazusa DNA Research Institute
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Characterization of Equine Microsatellites and Microsatellite-Linked Repetitive Elements (eMLREs) by Efficient Cloning and Genotyping Methods

Teruaki Tozaki1,2,*, Suguru Mashima2, Kei-ichi Hirota2, Nam-HO Choi-Miura1 and Motowo Tomita1

1Department of Physiological Chemistry, School of Pharmaceutical Sciences, Showa University Shinagawa,Tokyo 142-8555, Japan
2Department of Molecular Genetics, Laboratory of Racing Chemistry 1731-2 Tsuruta-cho, Utsunomiya, Tochigi 320-0851, Japan

* To whom correspondence should be addressed. Department of Molecular Genetics, Laboratory of Racing Chemistry, 1731-2 Tsuruta-cho, Utsunomiya, Tochigi 320-0851, Japan. Tel. +81-28-647-4472, Fax. +81-28-647-4473, E-mail: ttozaki{at}nyc.odn.ne.jp

We performed efficient cloning and genotyping methods for isolation of a large number of polymorphic microsatellites. The methods contain the time-efficient cloning method of constructing microsatellite-enriched libraries and the economic genotyping method of fluorescent labeling of PCR products. Eighty novel equine microsatellites cloned were efficiently isolated from the enrichment library and analyzed for genotype polymorphism. Of these, 72 microsatellites were analyzed with a good resolution. The average heterozygosity of all loci was 0.52, and the number of alleles ranged from one to 9 with an average of 4.5 alleles. The other eight loci showed multiple bands of PCR products, suggesting the occurrence of microsatellites in a repetitive element, in which the number of microsatellite repeats varies among different members of the repetitive element.

We found five homologous groups at flanking regions in comparison with the flanking regions of microsatellites from DNA databases. One of them showed homology to equine repetitive element-2. In the other four homologous groups, the two groups were named equine microsatellite-linked repetitive element-1 (eMLRE-1) and equine microsatellite-linked repetitive element-2 (eMLRE-2) as novel equine repetitive elements identified from equine genome. These data should help the analysis of equine DNA sequences and the design of equine genome markers.

Key words: Equine; microsatellite; repetitive element; polymorphism; MLRE


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 J HeredHome page
T. Tozaki, N. Takezaki, T. Hasegawa, N. Ishida, M. Kurosawa, M. Tomita, N. Saitou, and H. Mukoyama
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[Abstract] [Full Text] [PDF]


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